Reversing advanced glycosylation cross-links using heterocyclic-substituted thiazolium salts

ABSTRACT

The present invention relates to compositions and methods for reversing advanced glycosylation end product-mediated cross-linking and protein aging. Accordingly, compositions are described which comprise thiazolium compounds substituted with heterocyclic groups which are capable of reversing the formation of advanced glycosylation end product cross-links. Both industrial and therapeutic applications for the invention are disclosed, as food spoilage and animal protein aging can be treated. Such compounds have particular application in the treatment of protein aging such as is responsible for the complications of aging and diabetes.

BACKGROUND OF THE INVENTION

[0001] The present invention relates generally to the aging of proteinsresulting from their reaction with glucose and other reducing sugars,and more particularly to the reversing or cleavage of cross-links formedas a consequence of the formation of advanced glycosylation (glycation)end products.

[0002] The reaction between glucose and proteins has been known for sometime. Its earliest manifestation was in the appearance of brown pigmentsduring the cooking of food, which was identified by Maillard in 1912,who observed that glucose or other reducing sugars react with aminoacids to form adducts that undergo a series of dehydrations andrearrangements to form stable brown pigments. Further studies havesuggested that stored and heat treated foods undergo nonenzymaticbrowning as a result of the reaction between glucose and the polypeptidechain, and that the proteins are resultantly cross-linked andcorrespondingly exhibit decreased bioavailability. As described incopending application Ser. No. 08/588,249, incorporated herein byreference, these reactions have a parallel in vivo, and have been foundto occur with a variety of other body proteins, such as lenscrystallins, collagen and nerve proteins. These reactions areaccelerated in the presence of elevated glucose levels, as occur inindividuals with diabetes mellitus, but still occur in vivo at normalglucose levels. Termed advanced glycosylation (or glycation) endproducts (AGEs), the cross-linked products involving structural andother proteins within the body leads not only to aberrantphysico-chemical properties of, for example, connective tissue, but alsoresults in the formation of new chemical structures which are recognizedby specific receptors on various cell types and as a consequence oftheir recognition, initiate pathogenetic mechanisms leading to thecomplications of diabetes and aging.

[0003] Several successful therapeutic approaches have been achievedbased upon intervening in the accumulation of AGEs in vivo. Oneapproach, exemplified in U.S. Pat. No. 4,758,583, incorporated herein byreference, concerns the inhibition of the formation of AGEs from itsprecursors, by the administration of agents such as aminoguanidine andrelated compounds. By reacting with an early glycosylation product thatresults from the original reaction between the target protein andglucose, these agents block the formation of AGEs and further formationof AGEs and cross-links in tissues is inhibited. Efficacy of thisapproach has been demonstrated in numerous animal models of diabetes andaging, including positive effects on macrovascular, renal, retinal, andneural pathology. These data have been reviewed by Vlassara et al.,1994, “Biology of Diseases. Pathogenic effects of advancedglycosylation: biochemical, biologic and clinical implications fordiabetes and aging,” Laboratory Investigation 70:138-151; Brownlee,1995, “The pathological implications of protein glycation,” Clin.Invest. Med., 18:275-281; and Brownlee, 1995, “Advanced proteinglycosylation in diabetes and aging,” Ann. Rev. Med. 46:223-34.

[0004] In another pharmacological approach to controlling levels of AGEsin tissues, especially in those tissues in which AGE cross-links havealready accumulated to levels which are responsible for subclinical orclinical pathology, administration of agents that reverse or break AGEcross-links has proven successful. As described in U.S. Pat. No.5,656,261 and copending U.S. application Ser. Nos. 08/583,249 and08/848,776, all of which are incorporated herein by reference in theirentireties, agents and methods are disclosed which reverse (also termedcleave or break) existing AGE cross-links in vitro and in vivo. Studiesdemonstrate positive effects of such agents on cardiovascularcomplications related to aging which are accelerated in experimentaldiabetes (see Wolffenbuttel et al., 1998, “Breakers of AdvancedGlycation End Products Restores Large Artery Properties in ExperimentalDiabetes,” Proc. Nat. Acad. Sci. U.S.A. 95:4630-4634). In these studies,rats diabetic for 9 weeks followed by 1 to 3 weeks administration of anAGE breaker compound resulted in reversal of diabetes-induced increasesin large artery stiffness. Parameters that were improved includedcardiac output, peripheral resistance, systemic arterial compliance,input impedance of the aorta, and compliance of the carotid artery.

[0005] It is toward the identification of additional agents capable ofreversing AGE cross-links that the present invention is directed.

SUMMARY OF THE INVENTION

[0006] In accordance with the present invention, compounds and methodare disclosed for reversing AGE cross-linking. AGE cross-linking causedby other reactive sugars present in vivo or in foodstuffs, includingribose, galactose and fructose would also be reversed by the methods andcompositions of the present invention.

[0007] The agents useful in the present invention are members of theclass of compounds known as thiazoliums, and in particular thiazoliumcompounds substituted with heterocyclic groups.

[0008] The agents comprise compounds having the following structuralformula:

[0009] wherein

[0010] R¹ and R² are independently selected from the group consisting ofhydrogen and an alkyl group optionally substituted by a hydroxy group;

[0011] Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ring,members and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl; andsaid one or more substituents optionally substituted by one or morealkyl or alkoxy groups;

[0012] or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is aheterocyclic group containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen. nitrogen, and sulfur; saidheterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups;

[0013] and X is a halide, tosylate, methanesulfonate ormesitylenesulfonate ion.

[0014] The ability to reverse already-formed advanced glycosylationproducts carries with it significant implications in all applicationswhere advanced glycation and concomitant molecular crosslinking is aserious detriment. In the area of food technology, for instance, thecleavage of cross-links would confer a reversal of the increasedtoughness resulting from the formation of AGEs during storage. In apreferred embodiment, the application of agents capable of reversal ofthe Maillard process has particular benefit in vivo as AGE cross-linkingadversely affects several of the significant protein masses in the body,among them collagen, elastin, lens proteins, and the kidney glomerularbasement membrane. These proteins deteriorate both with age (hence theapplication of the term “protein aging”) and more rapidly as aconsequence of diabetes. Accordingly, the ability to reverse thecross-linking of these proteins and thus to reduce the amount ofcross-links present between advanced glycosylation end products andother proteins in the body carries the promise for treatment of thecomplications of diabetes and aging for instance, and thereby improvingthe quality and, perhaps, duration of animal and human life.

[0015] It is a yet further object of the present invention to provideagents which reverse the advanced glycosylation end products formed as aconsequence of the aforesaid advanced glycosylation reaction sequence bycleaving the α-dicarbonyl-based protein crosslinks present in theadvanced glycosylation end products.

[0016] It is a still further object of the present invention to providetherapeutic methods of treating the adverse consequences of molecular orprotein aging by resort to the aforesaid method and agents to achievethe reversal or cleavage of cross-links derived from advancedglycosylation reactions.

[0017] It is a still further object of the present invention to providecompositions, including pharmaceutical compositions, incorporating theagents of the present invention.

[0018] Other objects and advantages will become apparent to thoseskilled in the art from a consideration of the ensuing description.

DETAILED DESCRIPTION OF THE INVENTION

[0019] In accordance with the present invention, agents, compositionsincluding pharmaceutical compositions containing said agents andassociated methods are described which reverse already-formed advancedglycosylation end product-derived cross-links (AGE cross-links). Usefulagents, for instance, comprise compounds having, the structural formula:

[0020] wherein

[0021] R¹ and R² are independently selected from the group consisting ofhydrogen and an alkyl group optionally substituted by a hydroxy group;

[0022] Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl aryl, and aralkyl groups;and said one or more substituents optionally substituted by one or morealkyl or alkoxy groups;

[0023] or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is aheterocyclic group containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen, and sulfur; saidheterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups;

[0024] and X is a halide, tosylate, methanesulfonate ormesitylenesulfonate ion.

[0025] The heterocyclic groups referred to herein include 4-8 memberedrings having at least one and up to 3 heteroatoms, e.g., oxygen,nitrogen, or sulfur, therein; and including various degrees ofunsaturation. Representatives of such heterocyclic groups are those suchas isoxazolyl, phenylisoxazolyl, furanyl, morpholino, thiomorpholino,pyrimidinyl, piperidino, homopiperidino, piperazino, methylpiperazino,hexamethyleneimino, tetrahydroquinolyl, pyridyl, methylpyridyl,imidazolyl, pyrrolidinyl, 2,6-dimethylmorpholino, furfuryl,1,2,4-triazoylyl, thiazolyl, thiophenyl, thiazolinyl, methylthiazolyl,and the like. Excluded are alkylenedioxyaryl substituents, which aredescribed in copending application Ser. No. 08/588,249, incorporatedherein by reference. The heterocyclic groups of the present inventionmay be further substituted, for example, by an oxo group, to form, forexample, a 2-oxo-tetrahydroquinolinyl group, or substituted by one ormore alkyl, alkoxycarbonylalkyl, aryl, or aralkyl groups, and suchsubstituents may be further substituted by one or more alkyl or alkoxygroups.

[0026] Examples of Y groups of the compounds of the present include butare not limited to: 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-2-(1-piperidinyl)-2-oxoethyl);3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].

[0027] The alkyl groups referred to above contain one to about eighteencarbon atoms and include, for example, methyl, ethyl, propyl, butyl,pentyl, hexyl, octyl, decyl, dodecyl, and octadecyl, and thecorresponding branched-chain isomers thereof. Lower alkyl groups, of oneto about six carbon atoms, are preferred. The alkyl groups optionallysubstituted by hydroxy groups include alkyl groups as hereinbeforedefined substituted with a hydroxy group at any position, such as butnot limited to the following examples: hydroxymethyl, 1-hydroxyethyl,2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl,6-hydroxyhexyl, and the like. Hydroxy-substituted lower alkyl groups asdefined above are preferred. Similarly, the alkoxy groups contain fromone to about eighteen carbon atoms, and include, for example, methoxy,ethoxy, propoxy, butoxy, pentoxy, hexoxy, decoxy, and octadecoxy, andthe corresponding branched-chain isomers thereof. Lower alkoxy groups ofone to about 6 carbons, are preferred. The alkoxycarbonylalkyl groupsencompassed by the above formula include those wherein the alkoxyportion contain from one to about eighteen carbon atoms and the alkylportion contains from 1 to about eighteen carbon atoms. Typicalalkoxycarbonyl portions are those such as acetoxy or ethanoyloxy,propanoyloxy, butanoyloxy, pentanoyloxy, hexanoyloxy, decanoyloxy, andoctadecanoyloxy, and the corresponding branched chain isomers thereof.The preferred alkyl portions of these molecules have from one to aboutsix carbon atoms.

[0028] The aryl groups encompassed by the above formula are thosecontaining 6-10 carbon atoms, such as naphthyl, phenyl and alkyl oralkoxy substituted-phenyl, e.g., toluyl and xylyl.

[0029] For the purposes of this invention, the compounds of the presentinvention are formed as biologically and pharmaceutically acceptablesalts. Useful salt forms are the halides, particularly the bromide andchloride, tosylate, methanesulfonate, and mesitylenesulfonate salts.Other related salts can be formed using similarly non-toxic, andbiologically and pharmaceutically acceptable anions.

[0030] Of the compounds encompassed herein, certain substituents arepreferred. For instance, the compounds wherein R₁ or R₂ are hydrogen oralkyl groups are preferred. Also highly preferred are the compoundswherein Y is a 2-oxoethyl group with a heterocyclic group of thiophenyl,thiomorpholinyl, furanyl, 2-oxo-tetrahydroquinolinyl, and pyrrolidinyl.

[0031] As described in the formula above, the heterocyclic group may berepresented by the R group of the formula —CH₂C(═O)—R, or it mayrepresent the R′ group of the formula —CH₂C(═O)NHR′. Representative,non-limiting examples of compounds of the present invention are:

[0032] 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide

[0033] 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0034]3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide

[0035]3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethyl-thiazoliumchloride

[0036]3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(6-hydroxyhexyl)-thiazoliumbromide

[0037]3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0038] 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0039] 3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0040] 3-(2-(2-furanyl)-2-oxoethyl)-4-(2-hydroxypentyl)thiazoliumbromide

[0041]3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0042] 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide

[0043]3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0044] 3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0045] 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide

[0046] 3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazoliumbromide

[0047] 3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide

[0048] 3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide

[0049]3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0050] 3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide

[0051] 3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide

[0052] 3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0053]3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0054] 3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0055] 3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0056] 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-octylthiazoliumbromide

[0057] 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0058]3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dipropylthiazoliumchloride

[0059] 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dioctadecylthiazoliumbromide

[0060]3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dipentylthiazoliumbromide

[0061] 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-didodecylthiazolium bromide

[0062] 3-(2-(2-furanyl)-2-oxoethyl)-5-decylthiazolium bromide

[0063]3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dioctylthiazoliumbromide

[0064] 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-diethylthiazolium bromide

[0065]3-(2-(3-methyl-2-thianaphthenyl)-2oxoethyl]-4,5-dipentylthiazoliumbromide

[0066] 3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-thiazolium bromide

[0067] 3-(2-(2-thienyl)-2-oxoethyl)-thiazolium bromide

[0068]3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-(6-hydroxyhexyl)thiazoliumbromide

[0069] 3-(2-(4-thiomorpholinyl)-2-oxoethyl)thiazolium bromide

[0070] 3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dioctylthiazoliumbromide

[0071] 3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-didecylthiazoliumbromide

[0072] 3-(2-(2-pyridinyl )-2-oxoethyl)-4,5-dioctylthiazolium bromide

[0073] 3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dipropylthiazoliumchloride

[0074] 3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4-methylthiazoliumchloride

[0075] 3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-5-methylthiazoliumchloride

[0076] 3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4-octylthiazoliumchloride

[0077] The above compounds are capable of reversing already-formedadvanced glycosylation end products on proteins. The cross-linking ofproteins by formation of advanced glycosylation end products contributesto the entrapment of other proteins and results in the development invivo of conditions such as reduced elasticity and wrinkling of the skin,certain kidney diseases, atherosclerosis, osteoarthritis and the like.Similarly, foodstuffs including plant and animal material that undergoesnonenzymatic browning deteriorates and becomes spoiled or toughened and,consequently, inedible, unpalatable or non-nutritious. Thus, thecompounds employed in accordance with this invention reduce the level ofthe advanced glycosylation end product-associated cross-links alreadypresent in the protein material.

[0078] The present methods and compositions hold the promise forreversing the aging of key proteins both in animals and plants, andconcomitantly, conferring both economic and medical benefits as a resultthereof. In the instance of foodstuffs, the administration of thepresent composition holds the promise for reversing the physico-chemicalchanges imparted to foodstuffs on storage, such as the increasedtoughness of meats that occurs during aging or storage.

[0079] The therapeutic implications of the present invention relate tothe reversal of the aging process which has, as indicated earlier, beenidentified and exemplified in the aging of key proteins by advancedglycosylation and cross-linking. Thus, body proteins, and particularlystructural body proteins, such as collagen, elastin, lens proteins,nerve proteins, kidney glomerular basement membranes and otherextravascular matrix components would all benefit in their longevity andoperation from the practice of the present invention. The presentinvention thus reduces the incidence of pathologies involving theentrapment of proteins by cross-linked target proteins, such asretinopathy, cataracts, diabetic kidney disease, glomerulosclerosis,peripheral vascular disease, arteriosclerosis obliterans, peripheralneuropathy, stroke, hypertension, atherosclerosis, osteoarthritis,periarticular rigidity, loss of elasticity and wrinkling of skin,stiffening of joints, glomerulonephritis, etc. Likewise, all of theseconditions are in evidence and tend to occur at an accelerated rate inpatients afflicted with diabetes mellitus as a consequence of theirhyperglycemia. Thus, the present therapeutic method is relevant totreatment of these and related conditions in patients either of advancedage or those suffering from one of the mentioned pathologies.

[0080] Molecular cross-linking through advanced glycosylation productformation can decrease solubility of structural proteins such ascollagen in vessel walls and can also trap serum proteins, such aslipoproteins to the collagen. Also, the presence of advancedglycosylation end products and associated cross-links may result inincreased permeability of the endothelium and consequently covalenttrapping of extravasated plasma proteins in subendothelial matrix, aswell as a reduction in susceptibility of both plasma and matrix proteinsto physiologic degradation by enzymes. For these reasons, theprogressive occlusion of diabetic vessels induced by chronichyperglycemia has been hypothesized to result from excessive formationof sugar-derived and particularly, glucose-derived cross-links. Suchdiabetic microvascular changes and microvascular occlusion can beeffectively reversed by chemical reversal or cleavage of the advancedglycosylation product cross-link formation utilizing a composition andthe methods of the present invention.

[0081] Certain of the compounds disclosed herein are novel:

[0082] wherein

[0083] R¹ and R² are independently selected from the group consisting ofhydrogen and an alkyl group, optionally substituted by a hydroxy group;

[0084] Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and one heteroatom selected from the group consisting of sulfurand nitrogen, or 2-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl groups;and said one or more substituents optionally substituted by one or morealkyl or alkoxy groups;

[0085] or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is aheterocyclic group containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen, and sulfur; saidheterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups;

[0086] and X is a halide, tosylate, methanesulfonate ormesitylenesulfonate ion.

[0087] The heterocyclic groups referred to herein include 4-8 memberedrings having at least one nitrogen or sulfur atom, or 4-8 membered ringshaving 2-3 heteroatoms, e.g., oxygen, nitrogen, or sulfur, therein; andincluding various degrees of unsaturation. Representatives of suchheterocyclic groups are those such as isoxazolyl, phenylisoxazolyl,morpholino, thiomorpholino, pyrimidinyl, piperidino, homopiperidino,piperazino, methylpiperazino, hexamethyleneimino, tetrahydroquinolyl,pyridyl, methylpyridyl, imidazolyl, pyrrolidinyl,2,6-dimethylmorpholino, furfuryl, 1,2,4-triazoylyl, thiazolyl,thiophenyl, thiazolinyl, methylthiazolyl, and the like. Excluded arealkylenedioxyaryl substituents, which are described in co-pendingapplication Ser. No. 08/588,249, incorporated herein by reference. Theheterocyclic groups of the present invention may be further substituted,for example, by an oxo group, to form, for example, a2-oxo-tetrahydroquinolinyl group, or substituted by one or more alkyl,alkoxycarbonylalkyl, aryl, or aralkyl groups, and such substituents maybe further substituted by one or more alkyl or alkoxy groups.

[0088] The alkyl groups, alkoxy groups, alkoxycarbonylalkyl groups, arylgroups, and salts are as described hereinabove.

[0089] As described in the formula above, the heterocyclic group may berepresented by the R group of the formula —CH₂C(═O)—R, or it mayrepresent the R′ group of the formula —CH₂C(═O)NHR′. Representativeexamples of novel compounds of the present invention are:

[0090] 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]thiazolium bromide

[0091] 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0092]3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)thiazoliumbromide

[0093]3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0094] 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0095]3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0096] 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0097]3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0098] 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0099]3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0100] 3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide

[0101] 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0102] 3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazoliumbromide

[0103] 3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide

[0104] 3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide

[0105]3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0106] 3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide

[0107] 3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0108] 3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0109]3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0110] 3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0111] 3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride

[0112] As described in the Background section above, studies on otherAGE cross-link cleavage agents have shown the ability to reversecardiovascular damage in an experimental model of diabetes. As diabetesis often considered to be a model of accelerated aging, especially withrespect to macrovascular complications, the agents of the presentinvention are suitable for therapeutic use in such complications ofaging, examples of which are described above.

[0113] As will be shown in the Examples below, and as cited above,compounds capable of reversing AGE cross-links show significant promisein the treatment of various diseases and complications related toprotein aging. Compounds active in an in vitro model of AGEcross-linking in which the compounds are evaluated for their ability toreverse the covalent cross-linking of IgG to red blood cell membranes,or cleave collage cross-links, has corresponding in-vivo activity in amodel of protein aging. As described above, such studies demonstratepositive effects of such agents on cardiovascular complications relatedto aging which are accelerated in experimental diabetes (seeWolffenbuttel et al., 1998. “Breakers of Advanced Glycation End ProductsRestores Large Artery Properties in Experimental Diabetes,” Proc. Nat.Acad. Sci. U.S.A. 95:4630-4634). In these studies, rats diabetic for 9weeks followed by 1 to 3 weeks administration of an AGE breaker compoundresulted in reversal of diabetes-induced increases in large arterystiffness. Parameters that were improved included cardiac output,peripheral resistance, systemic arterial compliance, input impedance ofthe aorta, and compliance of the carotid artery. Thus, activeAGE-reversing compounds of the present invention are candidates for thetreatment of the complications of protein aging.

[0114] In the instance where the compositions of the present inventionare utilized for in vivo or therapeutic purposes, it may be noted thatthe compounds or agents used therein are biocompatible. Variousbiologically or pharmaceutically acceptable salts may be used, such ashalides, tosylate, etc. Pharmaceutical compositions may be prepared witha therapeutically effective quantity of the agents or compounds of thepresent invention and may include a pharmaceutically acceptable carrier,selected from known materials utilized for this purpose. Suchcompositions may be prepared in a variety of forms, depending on themethod of administration. Also, various pharmaceutically acceptableaddition salts of the compounds of the present invention may beutilized.

[0115] A liquid form would be utilized in the instance whereadministration is by intravenous, intramuscular or intraperitonealinjection. When appropriate, solid dosage forms such as tablets,capsules, or liquid dosage formulations such as solutions andsuspensions, etc., may be prepared for oral administration. For topicalor dermal application to the skin or eye, a solution, a lotion orointment may be formulated with the agent in a suitable vehicle such aswater, ethanol, propylene glycol, perhaps including a carrier to aid inpenetration into the skin or eye. For example, a topical preparationcould include up to about 10% of the compound of Formula I. Othersuitable forms for administration to other body tissues are alsocontemplated.

[0116] In the instance where the present method has therapeuticapplication, the animal host intended for treatment may haveadministered to it a quantity of one or more of the agents, in asuitable pharmaceutical form. Administration may be accomplished byknown techniques, such as oral, topical and parenteral techniques suchas intradermal, subcutaneous, intravenous or intraperitoneal injection,as well as by other conventional means. Administration of the agents maytake place over an extended period of time at a dosage level of, forexample, up to about 30 mg/kg.

[0117] The agent of the present invention is formulated in compositionsin an amount effective to inhibit and reverse the formation of advancedglycosylation end products. This amount will, of course, vary with theparticular agent being utilized and the particular dosage form, buttypically is in the range of 0.01% to 1.0%, by weight, of the particularformulation.

[0118] The compounds of the present invention can be prepared generallyaccording to the methods described in Potts et al., 1976. J. Org. Chem.41:187, and Potts et al., 1977, J. Org. Chem. 42:1648, or as shown inthe following, scheme wherein R is a heterocyclic group, R¹, R², and Zare as described hereinabove, and X is a halogen atom:

[0119] In the reaction scheme above, the appropriate substitutedthiazole compounds of formula I is reacted with the appropriate halocompound of formula II, to afford the desired compound of the presentinvention; all substituents are as hereinbefore defined.

[0120] The halo reactant may be prepared by suitable techniques known inthe art. For example, for the preparation of3-(2-thiophenyl-2-oxoethyl)-4,5-dimethyl-thiazolium bromide, thereactant 2-bromothiophene is reacted with dimethylthiazole.2-Bromothiophene may be prepared according to the method of King, etal., 1964, J. Org. Chem. 29:3459, by the bromination of2-acetylthiophene with copper (II) bromide. Specific methods aredescribed in the examples below.

[0121] The conditions for the reaction between the halo compound and thethiazole derivative generally involve refluxing the mixture at 110° C.in an oil bath for 3-7 hours with a minimum amount of solvent such asacetonitrile, or refluxing the mixture in ethanol or acetonitrile for3-5 hours. If the halo reactant contains chlorine, the first conditionis used. For a bromo compound, the second condition is preferable.

[0122] The present invention may be better understood by reference tothe following non-limiting Examples, which are provided as exemplary ofthe invention. The following examples are presented in order to morefully illustrate the preferred embodiments of the invention. They shouldin no way be construed, however, as limiting the broad scope of theinvention.

EXAMPLE 1 Preparation of3-(2-(1-thienyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide

[0123] This compound was prepared by reacting 4,5-dimethylthiazole with2-bromoacetylthiophene. 2-Bromoacetylthiophene was prepared bybromination of 2-acetylthiophene with copper (II) bromide according tothe method of King et al., 1964, J. Org. Chem. 29:3459.2-Acetylthiophene (6 g, 47.6 mmol) was dissolved in chloroform (60 mL)and added to a slurry of copper (II) bromide (13.5 g, 60.44 mmol) inethyl acetate (120 mL). The mixture was refluxed for 6 hours and thenfiltered while still hot through a celite pad. The filtrate cake waswashed with ethyl acetate and the combined filtrate was evaporated togive 2-bromoacetylthiophene (9.5 g, 97.3%). The crude product was useddirectly in the next reaction.

[0124] A solution of 4,5-dimethylthiazole (2.2 g, 19.4 mmol) and2-bromoacetylthiophene (4 g, 19.4 mmol) in ethyl alcohol (20 mL) wasrefluxed for 3 hours. It was cooled to room temperature and t-butylmethyl ether (10 mL) was added. The reaction mixture was left at roomtemperature overnight with stirring. The white product separated and wasfiltered and dried. It was crystallized from ethyl alcohol (4.42 g.73%). m.p. 203-205° C.

EXAMPLE 2 Synthesis of3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethylthiazolium chloride

[0125] 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumchloride was prepared from the reaction of N-(chloroacetyl)pyrrolidinewith 4,5-dimethylthiazole. N-(chloroacetyl)pyrrolidine was prepared asfollows. Pyrrolidine (63.9 g, 0.9 mol) was taken in methylene chloride(640 mL) and cooled to 0° C. in a salt-ice bath. To the stirred mixturewas added chloroacetyl chloride (101.8 g in 450 mL of CH₂Cl₂, 0.9 mol)dropwise keeping the inside temperature below 15° C. After adding thechloroacetyl chloride,, the mixture was stirred for an hour at 5° C.Sodium hydroxide solution (7 M, 190 mL) was added with vigorous stirringsuch that the inside temperature did not exceed 20° C. The mixture wasstirred for 15 minutes and the aqueous layer was separated. The organiclayer was washed successively with saturated sodium bicarbonatesolutions (2×200 mL), water (1×200 mL) and dried over anhydrous sodiumsulfate. The solvent was removed in vacuo and the residue wasrecrystallized from hexane to give 64.5 g (48.6%) of white platecrystals, m.p. 43° C.

[0126] A mixture of N-(chloroacetyl)pyrrolidine (37.3 g, 0.25 mol),4,5-dimethylthiazole (28.69 g, 0.25 mol) and acetonitrile (17 mL) wasrefluxed in an oil bath at 105-110° C. for 7 hours. To the reactionmixture was added acetonitrile (230 mL) and continued to heat for 20minutes, then t-butyl methyl ether (250 mL) was added. The reactionmixture was kept at room temperature overnight. The product was filteredand washed with a mixture of t-butyl methyl ether and acetonitrile (1:1v/v, 100 mL), and t-butyl methyl ether (150 mL) to obtain 59.64 g (90.2%yield) of white solid. The crude product (59.64 g) was dissolved inacetonitrile (350 mL) with heating, filtered, t-butyl methyl ether (350mL) added, and the solution was allowed to cool at room temperature for3 hours. The product was filtered and washed with a mixture of t-butylmethyl ether and acetonitrile (1:1 v/v, 300 mL), and t-butyl methylether (300 mL) to yield 56.95 g (86.1% yield) of white crystals, m.p.196-198° C.

EXAMPLE 3

[0127] Using the procedures described above in Examples 1 and 2, thefollowing compounds were prepared using the corresponding reactants. Themelting points are indicated.3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide 213-214° C.3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide245-246° C. (dec.)3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide 209-210° C. (dec.)3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethyl-thiazolium120-121° C. chloride3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide 197-198°C. (dec.)3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide 215-216° C.3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide 240-242°C. 3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide 195-196°C. (dec.)3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide 274-275° C. (dec.)3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium chloride189-198° C.3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide 171-172° C.3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide150-152° C. 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide203-205° C.3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide175-176° C. 3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumchloride 232-233° C. (dec.)3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium chloride218-219° C. (dec.)3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 215-216° C. (dec.)3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium chloride210-212° C. (dec.)3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide 178-179°C. 3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 220-221° C. (dec.)3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 248-249° C. (dec.)3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 170-171° C.3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 180-182° C.

EXAMPLE 4

[0128] A typical pharmaceutical dosage form is prepared as follows.mg/tablet Compound of the present invention 50 Starch 50 Mannitol 75Magnesium stearate 2 Stearic acid 5

[0129] The compound, a portion of the starch and the lactose arecombined and wet granulated with starch paste. The wet granulation isplaced on trays and allowed to dry overnight at a temperature of 45° C.The dried granulation is comminuted in a comminutor to a particle sizeof approximately 20 mesh. Magnesium stearate, stearic acid and thebalance of the starch are added and the entire mix blended prior tocompression on a suitable tablet press. The tablets are compressed at aweight of 232 mg. using a {fraction (11/32)}″ punch with a hardness of 4kg. These tablets will disintegrate within a half hour according to themethod described in USP XVI.

EXAMPLE 5

[0130] Lotion mg/g Compound of Formula I 1.0 Ethyl alcohol 400.0Polyethylene glycol 400 300.0 Hydroxypropyl cellulose 5.0 Propyleneglycol to make 1.0 g

EXAMPLE 6 Reversing Age Cross-links

[0131] The activity of the compounds of the present invention to reverseAGE cross-links was evaluated in an in-vitro model, as follows. Bloodwas collected from streptozotocin-diabetic rats (6 to 8 wk diabetic) inheparinized tubes, centrifuged at 2500×g for 15 min. at 40° C., and theplasma and buffy-coat layers aspirated off. The RBC were washed with PBS(ca. 5 ml PBS per ml blood) three times.

[0132] Test compounds were dissolved in PBS and the pH adjusted to 7.0,if necessary. Two hundred microliters of washed RBC were added to 2 mLof test compound solution, and the mixture was incubated overnight at37° C. For a control, 200 μl of RBC was incubated in 2 mL of PBS.

[0133] After the overnight incubation, the reaction mixtures werecentrifuged and the RBC pellets were washed three times with PBS,followed by diluting 1:30 in PBS. Immunoglobulin G (IgG) bound to thesurface of the washed RBCs was then determined.

[0134] Assay for RBC-IgG: The assay is performed in a Multiscreen-HA.0.45 μm cellulose mixed esters membrane-sealed 96 well plate (MilliporeMAHAS45 ). The membranes are first wet by filling the wells with 100 uLPBS, and the wells emptied by applying a vacuum to the Millititer vacuummanifold. Three hundred uL of Superblock (pre-warmed to 37° C.) is thenadded to each well and incubated at 37° C. for one hour. The Superblockis then removed from the wells by application of vacuum, then the wellsare washed once with 300 ul of PBS-Tween (0.05%) and three times withPBS. The vacuum is then turned off.

[0135] One hundred μl of PBS is then added to each well. Each RBC sampleis gently vortexed, and 50 uL pipetted into the wells, in sextuplicate.The wells labeled A1-A3 are left for a reagent blank and wells A4-A6reserved for an antibody blank. Vacuum is then applied to remove thebuffer and the RBCs are washed once with PBS. Alkalinephosphatase-labeled rabbit-anti-rat IgG is diluted 1 to 25000 in PBS,and 50 μl is added to triplicate wells of each sample. PBS alone isadded to the other three wells of each sample. These will serve assample blanks to adjust for any endogenous alkaline phosphataseactivity. The samples are allowed to stand at room temperature for twohours. The solution is then removed by application of vacuum and the redblood cells are washed with PBS-Tween twice, PBS twice and with TBStwice. The bottom of the plate is rinsed with distilled water andblotted dry with paper towels. To each well is then added 100 μl ofp-nitrophenyl phosphate substrate (1 mg/ml in diethanolamine buffer pH9.5), and the color is allowed to develop for two hours at 37° C. Thesolutions in each well are then transferred to a 96 well microtiterplate inside the vacuum manifold. One hundred μl of PBS to each well andvacuum applied again to transfer any remaining solution into themicrotiter plate. The OD at 410 nm is then read in a Dynatech Platereader (Sample filter 1 and Ref. filter 4).

[0136] For data analysis, breaking activity (percent reversal) isexpressed as the percent decrease in O.D. caused by incubation of RBCwith the test compound compared to RBC incubated in PBS alone. Using theabove assay, the following data was generated on the compounds of thepresent invention: Percent Compound reversal3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide24% 3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide 27%3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide 37%3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide 38% 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide38% 3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide47% 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide 53%3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide 63%3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide 76%3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride 70%3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazolium84% bromide

EXAMPLE 7 Reversing Collagen Cross-links

[0137] In a further evaluation of the ability of compounds of thepresent invention to reverse AGE cross-links between a circulatingprotein and collagen. AGE-BSA was cross-linked to rat tail tendoncollagen (type 1)-coated 96 well microtiter plates, and reversal ofcross-linking was determined.

[0138] AGE-BSA preparation. AGE-BSA is prepared by incubation of asolution of BSA (400 mg/ml) in 0.4 M sodium phosphate buffer pH 7.4 withan equal volume of 400 mM glucose solution in the same buffer at 37° C.for 12 weeks. The pH of the incubation mixture is monitored weekly andadjusted to pH 7.4 if necessary. After 12 weeks, the AGE-BSA solution isdialyzed against PBS for 48 hours with 4 buffer changes. Proteinconcentration is determined by the micro-Lowry method.

[0139] Assay protocol. A Biocoat (Collaborative Biomedical Products)plate is blocked with Superblock (300 mL /well) at 37° C. for 1 hr andwashed with PBS-Tween three times. AGE-BSA was diluted in PBS to aconcentration required to obtain maximum cross-linking as determined ina preliminary experiment. One hundred μl of the AGE-BSA working solutionis added to test wells and a similar concentration of BSA is added toblank wells. The first three wells are left empty for the reagent blank.The plate is incubated at 37° C. for four hours and washed withPBS-Tween three times. Test compounds are dissolved in PBS and pHadjusted to 7.0 if necessary. One hundred μl of a test compound is addedto triplicate wells. To measure maximum crosslinking 100 μl of PBS isadded to three to six wells. The test compounds and PBS are also addedto triplicate wells incubated with BSA, to obtain the blank readings.The plate is then incubated overnight at 37° C. The plate is washed withPBS-Tween, 50 mL of Rabbit-anti-BSA antibody (1 to 4000 in PBS) is addedto each well and the plate is incubated at room temperature for 60 min.After the plate is washed with PBS-Tween, 50 mL of horseradishperoxidase-conjugated-goat-anti-rabbit IgG (1 to 4000 in PBS) is addedto each well except the first three wells. The plate is incubated atroom temperature for 30 min. and washed with PBS-Tween. Two hundred μlof ABTS substrate (prepared from HRP-substrate buffer 10× (Sigma)diluted 1:10 in deionized water and mixed with 50×ABTS reagent (Sigma))is added to each well and color developed at 37° C. for 15 min.

[0140] Optical density is read at 410 nm with the sample filter set to“1” and the reference filter set to “5” on the Dynatech ELISA platereader.

[0141] Data Analysis: The average optical density (OD) is calculated foreach triplicate determination: Corrected OD=(Average of OD AGE-BSAwells−Average of OD BSA wells). Percent breaking by test compounds isexpressed as the percent decrease in OD of TTC-AGE-BSA wells incubatedwith test compounds compared to TTC-AGE-BSA wells incubated with PBS.

[0142] Using the procedure above, it was found that 3 mM3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide caused a 15%reversal of AGE cross-links.

[0143] This invention may be embodied in other forms or carried out inother ways without departing from the spirit or essentialcharacteristics thereof. The present disclosure is therefore to beconsidered as in all respects illustrative and not restrictive, thescope of the invention being indicated by the appended claims, and allchanges which come within the meaning and range of equivalency areintended to be embraced therein.

[0144] Various citations to the literature are presented herein, all ofwhich are incorporated herein in their entireties.

What is claimed is:
 1. A composition for reversing pre-formed advancedglycosylation crosslinks comprising an effective amount of a compound ofthe structural formula:

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl groups;and said one or more substituents optionally substituted by one or morealkyl or alkoxy groups; or a group of the formula —CH₂C(═O)—NHR′ whereinR′ is a heterocyclic group containing 4-10 ring members and 1-3heteroatoms selected from the group consisting of oxygen, nitrogen, andsulfur; said heterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups; and X is a halide, tosylate,methanesulfonate or mesitylenesulfonate ion; and a carrier therefor. 2.The composition of claim 1 wherein Y is a group of the formula—CH₂C(═O)R wherein R is a heterocyclic group other thanalkylenedioxyaryl containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur; saidheterocyclic group optionally substituted by one or more substituentsselected from the group consisting of alkyl, oxo, alkoxycarbonylalkyl,aryl, and aralkyl groups; and said one or more substituents optionallysubstituted by one or more alkyl or alkoxy groups; and X is a halide,tosylate, methanesulfonate or mesitylenesulfonate ion.
 3. Thecomposition of claim 2 wherein Y is selected from the group consistingof 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl); 3-(2-(2-furanyl)-2-oxoethyl;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].
 4. The composition of claim 3wherein said compound is3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide; 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide;3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide;3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride; or another biologically acceptable salt thereof.
 5. Thecomposition of claim 1 wherein said compound has the formula wherein Yis a group of the formula —CH₂C(═O)—NHR′ wherein R′ is a heterocyclicgroup containing 4-10 ring members and 1-3 heteroatoms selected from thegroup consisting of oxygen, nitrogen, and sulfur; said heterocyclicgroup optionally substituted by one or more alkoxycarbonylalkyl groups;and X is a halide, tosylate, methanesulfonate or mesitylenesulfonateion.
 6. The composition of claim 5 wherein said compound is3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethylthiazoliumchloride or another biologically acceptable salt thereof.
 7. Apharmaceutical composition for administration to an animal to reversepre-formed advanced glycosylation crosslinks within said animal,comprising a pharmaceutically effective amount of a compound selectedfrom the group consisting of compounds of the formula:

wherein R¹ and R² are independently selected from the group consistingof hydrogen and a alkyl group optionally substituted by a hydroxy group;Y is a group of the formula —CH₂C(═O)R wherein R is a heterocyclic groupother than alkylenedioxyaryl containing 4-10 ring members and 1-3heteroatoms selected from the group consisting of oxygen, nitrogen andsulfur; said heterocyclic group optionally substituted by one or moresubstituents selected from the group consisting of alkyl, oxoalkoxycarbonylalkyl, aryl, and aralkyl groups; and said one or moresubstituents optionally substituted by one or more alkyl or alkoxygroups; or a group of the formula —CH₂C(═O)—NHR′ wherein R′ is aheterocyclic group containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen, and sulfur; saidheterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups; and X is a halide, tosylate,methanesulfonate or mesitylenesulfonate ion, and a carrier therefor. 8.The composition of claim 7 wherein Y is a group of the formula—CH₂C(═O)R wherein R is a heterocyclic group other thanalkylenedioxyaryl containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur; saidheterocyclic group optionally substituted by one or more substituentsselected from the group consisting of alkyl, oxo, alkoxycarbonylalkyl,aryl, and aralkyl groups; and said one or more substituents optionallysubstituted by one or more alkyl or alkoxy groups; and X is a halide,tosylate, methanesulfonate or mesitylenesulfonate ion.
 9. Thecomposition of claim 8 wherein Y is selected from the group consistingof 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl); 3-(2-(2-furanyl)-2-oxoethyl;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].
 10. The composition of claim9 wherein said compound is3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide; 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide; 3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide;3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide;3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride; or another biologically acceptable salt thereof.
 11. Thecomposition of claim 7 wherein said compound has the formula wherein Yis group of the formula —CH₂C(═O)—NHR′ and R′ is a heterocyclic groupcontaining 4-10 ring members and 1-3 heteroatoms selected from the groupconsisting of oxygen, nitrogen, and sulfur; said heterocyclic groupoptionally substituted by one or more alkoxycarbonylalkyl groups; and Xis a halide, tosylate, methanesulfonate or mesitylenesulfonate ion. 12.The composition of claim 11 wherein said compound is3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethyl-thiazoliumchloride or another biologically acceptable salt thereof.
 13. A methodfor reversing pre-formed advanced glycosylation crosslinks, comprisingcontacting the target protein with an effective amount of compositioncomprising a compound selected from the group consisting of compounds ofthe formula

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkyl groups;and said one or more substituents optionally substituted by one or morealkyl or alkoxy groups; or a group of the formula —CH₂C(═O)—NHR′ whereinR′ is a heterocyclic group containing 4-10 ring members and 1-3heteroatoms selected from the group consisting of oxygen, nitrogen, andsulfur; said heterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups; and X is a halide, tosylate,methanesulfonate or mesitylenesulfonate ion; and a carrier therefor. 14.The composition of claim 13 wherein Y is a group of the formula—CH₂C(═O)R wherein R is a heterocyclic group other thanalkylenedioxyaryl containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur; saidheterocyclic group optionally substituted by one or more substituentsselected from the group consisting of alkyl, oxo, alkoxycarbonylalkyl,aryl, and aralkyl groups; and said one or more substituents optionallysubstituted by one or more alkyl or alkoxy groups; and X is a halide,tosylate, methanesulfonate or mesitylenesulfonate ion.
 15. Thecomposition of claim 14 wherein Y is selected from the group consistingof 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl); 3-(2-(2-furanyl)-2-oxoethyl;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].
 16. The composition of claim15 wherein said compound is3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide; 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide; 3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(-1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide;3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2- (2-thienyl)-2-oxoethyl)-4-dimethyl-5-hydroxyethylthiazoliumbromide; 3-(2-(4thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide; 3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride; or another biologically acceptable salt thereof.
 17. Thecomposition of claim 13 wherein said compound has the formula wherein Yis a group of the formula —CH₂C(═O)—NHR′ wherein R′ is a heterocyclicgroup containing 4-10 ring members and 1-3 heteroatoms selected from thegroup consisting of oxygen, nitrogen, and sulfur; said heterocyclicgroup optionally substituted by one or more alkoxycarbonylalkyl groups;and X is a halide, tosylate, methanesulfonate or mesitylenesulfonateion.
 18. The composition of claim 17 wherein said compound is3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethyl-thiazoliumchloride or another biologically acceptable salt thereof.
 19. A methodfor treating an animal to reverse pre-formed advanced glycosylationcrosslinks of a target protein within said animal, said methodcomprising administering an effective amount of a pharmaceuticalcomposition, said pharmaceutical composition comprising a compoundselected from the group consisting of compounds of the formula

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and 1-3 heteroatoms selected from the group consisting ofoxygen, nitrogen and sulfur; said heterocyclic group optionallysubstituted by one or more substituents selected from the groupconsisting of alkyl, oxo, alkoxycarbonylalkyl aryl, and aralkyl groups;and said one or more substituents optionally substituted by one or morealkyl or alkoxy groups; or a group of the formula —CH₂C(═O)—NHR′ whereinR′ is a heterocyclic group containing 4-10 ring members and 1-3heteroatoms selected from the group consisting of oxygen, nitrogen, andsulfur; said heterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups; and X is a halide, tosylate,methanesulfonate or mesitylenesulfonate ion; and a carrier therefor. 20.The composition of claim 19 wherein Y is a group of the formula—CH₂C(═O)R wherein R is a heterocyclic group other thanalkylenedioxyaryl containing 4-10 ring members and 1-3 heteroatomsselected from the group consisting of oxygen, nitrogen and sulfur; saidheterocyclic group optionally substituted by one or more substituentsselected from the group consisting of alkyl, oxo, alkoxycarbonylalkyl,aryl, and aralkyl groups; and said one or more substituents optionallysubstituted by one or more alkyl or alkoxy groups; and X is a halide,tosylate, methanesulfonate or mesitylenesulfonate ion.
 21. Thecomposition of claim 20 wherein Y is selected from the group consistingof 3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl); 3-(2-(2-furanyl)-2-oxoethyl;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].
 22. The composition of claim21 wherein said compound is3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide; 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide; 3-(2-(2-furanyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl-]-4,5-dimethylthiazoliumbromide; 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide;3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide;3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride; or another biologically acceptable salt thereof.
 23. Thecomposition of claim 19 wherein said compound has the formula wherein Yis a group of the formula —CH₂C(═O)—NHR′ wherein R′ is a heterocyclicgroup containing 4-10 ring members and 1-3 heteroatoms selected from thegroup consisting of oxygen, nitrogen, and sulfur; said heterocyclicgroup optionally substituted by one or more alkoxycarbonylalkyl groups;and X is a halide, tosylate, methanesulfonate or mesitylenesulfonateion.
 24. The composition of claim 23 wherein said compound is3-[2-[4-(2-ethoxy-2-oxoethyl)-2-thiazolyl]amino-2-oxoethyl]-4,5-dimethyl-thiazoliumchloride or another biologically acceptable salt thereof.
 25. A compoundof the formula

wherein R¹ and R² are independently selected from the group consistingof hydrogen and an alkyl group optionally substituted by a hydroxygroup; Y is a group of the formula —CH₂C(═O)R wherein R is aheterocyclic group other than alkylenedioxyaryl containing 4-10 ringmembers and one heteroatom selected from the group consisting ofnitrogen and sulfur, or 2-3 heteroatoms selected from the groupconsisting of oxygen, nitrogen and sulfur; said heterocyclic groupoptionally substituted by one or more substituents selected from thegroup consisting of alkyl, oxo, alkoxycarbonylalkyl, aryl, and aralkylgroups; and said one or more substituents optionally substituted by oneor more alkyl or alkoxy groups; or a group of the formula —CH₂C(═O)—NHR′wherein R′ is a heterocyclic group containing 4-10 ring members and 1-3heteroatoms selected from the group consisting of oxygen. nitrogen, andsulfur; said heterocyclic group optionally substituted by one or morealkoxycarbonylalkyl groups; and X is a halide, tosylate,methanesulfonate or mesitylenesulfonate ion.
 26. The compound of claim25 wherein said compound has the formula wherein Y is group of theformula —CH₂C(═O)R wherein R is a heterocyclic group other thanalkylenedioxyaryl containing 4-10 ring members and one heteroatomselected from the group consisting of nitrogen and sulfur, or 2-3heteroatoms selected from the group consisting of oxygen, nitrogen andsulfur; said heterocyclic group optionally substituted by one or moresubstituents selected from the group consisting of alkyl, oxo,alkoxycarbonylalkyl, aryl, and aralkyl groups; and said one or moresubstituents optionally substituted by one or more alkyl or alkoxygroups; and X is a halide, tosylate, methanesulfonate ormesitylenesulfonate ion.
 27. The composition of claim 26 wherein Y isselected from the group consistingof3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl];3-(2-(4-morpholinyl)-2-oxoethyl);3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl];3-(2-(1-piperidinyl)-2-oxoethyl);3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl];3-(2-(1-pyrrolidinyl)-2-oxoethyl;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl];3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl; 3-(2-(2-thienyl)-2-oxoethyl);3-(2-(2-thienyl)-2-oxoethyl); 3-(2-(4-thiomorpholinyl)-2-oxoethyl);3-(2-(hexahydro-1-azepinyl)-2-oxoethyl);3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl;3-(2-(octahydro-1-azocinyl)-2-oxoethyl); 3-(2-(2-pyridinyl)-2-oxoethyl;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl];3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]; and3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl].
 28. The composition of claim27 wherein said compound is3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-thiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4,5-dimethylthiazolium bromide;3-[2-(3-phenyl-5-isoxazolyl)-2-oxoethyl]-4-methyl-5-(2-hydroxyethyl)-thiazoliumbromide; 3-(2-(4-morpholinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2,6-dimethyl-4-morpholinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-piperidinyl)-2-oxoethyl)-4,5-dimethylthiazoliumbromide;3-[2-(2-oxo-1,2,3,4-tetrahydro-6-quinolinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(1-pyrrolidinyl)-2-oxoethyl)-4,5-dimethyl-thiazoliumbromide;3-[2-(3-methyl-2-thianaphthenyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide;3-[2-(4-phenyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumbromide; 3-(2-(2-thienyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-thienyl)-2-oxoethyl)-4-methyl-5-hydroxyethylthiazolium bromide;3-(2-(4-thiomorpholinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-(2-(hexahydro-1-azepinyl)-2-oxoethyl)-4,5-dimethylthiazolium bromide;3-[2-(4-[2-methoxyphenyl]-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-(2-(octahydro-1-azocinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-(2-(2-pyridinyl)-2-oxoethyl)-4,5-dimethyl-thiazolium bromide;3-[2-(2-methyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(2,6-dimethyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperidinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride;3-[2-(4-benzyl-1-piperazinyl)-2-oxoethyl]-4,5-dimethylthiazoliumchloride; or another biologically acceptable salt thereof.
 29. Thecomposition of claim 25 wherein said compound has the formula wherein Yis group of the formula —CH₂C(═O)—NHR′ and R′ is a heterocyclic groupcontaining 4-10 ring members and 1-3 heteroatoms selected from the groupconsisting of oxygen, nitrogen, and sulfur; said heterocyclic groupoptionally substituted by one or more alkoxycarbonylalkyl groups; and Xis a halide, tosylate, methanesulfonate or mesitylenesulfonate ion. 30.The composition of claim 29 wherein said compound is3-[2-(2′-amino-5′-carboethoxymethylene-thiazolyl)-2-oxoethyl]-4,5-dimethyl-thiazoliumchloride or another biologically acceptable salt thereof.